Finding the rare mutant allele needle in the wild-type haystack

Studies of biomarkers associated with the onset of and progression of cancers have been fraught with poor selectivity and have failed to detect mutant sequences with abundances of less than 1 in 100 wild-type sequences. The QX100 ddPCR platform offers a simple and robust means to increase the precision and sensitivity of TaqMan assays for rare event detections. Not only are direct counts of template molecules within a sample possible (Pinheiro et al. 2012), but partitioning mitigates the effects of target competition, greatly improving the discriminatory capacity of assays that differ by only a single nucleotide. Here, the benefits for clinically relevant assay targets are demonstrated through the BRAF V600E mutation assay, where the LOD improved >100-fold when performed in ddPCR. This platform should prove invaluable to the molecular diagnostics field in general and to the cancer field in particular for the routine assessment of mutation status. Combined with the high accuracy achieved through the use of tens of thousands of partitions, ddPCR represents a unique platform for extremely precise and sensitive measurements of target molecules.

Click here to learn more on how Bio-Rad Laboratories’ QX100™ Droplet Digital™ PCR System can aid in the detection of rare mutant alleles within a background of wild-type sequences.

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